Increased proliferation in keloid fibroblasts wounded in vitro
Increased proliferation in keloid fibroblasts wounded in vitro
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DOI：
10.1006/jsre.1996.0127
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发表时间：
1996-03-01
影响因子：
2.2
通讯作者：
Banes, AJ
中科院分区：
文献类型：
--
作者：
Calderon, M;Lawrence, WT;Banes, AJ
疤痕疙瘩是一种病理性过度生长的疤痕，超出了起始皮肤伤口的范围。最终，这种扩张性疤痕是伤口内成纤维细胞合成过量胶原蛋白的结果。导致这种胶原蛋白过量的过程仍然未知。开发了体外伤口模型来测试以下假设：从瘢痕疙瘩组织中分离并在体外受伤的成纤维细胞可能比类似受伤的正常真皮成纤维细胞增殖更快，瘢痕疙瘩成纤维细胞（KF）和正常人真皮成纤维细胞（NDF）在柔性底培养板中生长至汇合并静止，使用专门设计的夹具以标准化方式创建伤口，该夹具使用25号针来可重复地从汇合细胞片中消融 16-20% 的细胞。在受伤后24、48、72和96小时用H-3-胸苷标记受伤和未受伤的细胞以测量DNA合成。与未受伤的对照培养物相比，受伤的KF和NDF表现出H-3-胸苷掺入增加，并且受伤后24小时和48小时，受伤的KF表现出比受伤的NDF显着更高的H-3-胸苷掺入水平。细胞计数也出现了类似的趋势。通过放射自显影定量，受伤的 KF 还显示出统计上比受伤的 NDF 更高的标记指数。体外瘢痕疙瘩成纤维细胞中响应损伤而增加的 DNA 合成承诺与体内观察到的病理相关。与正常成纤维细胞相比，瘢痕疙瘩成纤维细胞进入S期的固有阈值可能较低，这导致瘢痕疙瘩成纤维细胞响应体外受伤而增殖增加。 (C) 1996 学术出版社
A keloid is a pathological overgrowth of scar expanding beyond the boundaries of the initiating skin wound. Ultimately, this expansive scar is a result of excess collagen synthesized by fibroblasts within the wound, The processes that lead to this collagen excess remain unknown. An in vitro wound model was developed to test the hypothesis that fibroblasts isolated from keloid tissue and wounded in vitro might proliferate more rapidly than similarly wounded normal dermal fibroblasts, Keloid fibroblasts (KF) and normal human dermal fibroblasts (NDF) were grown to confluence and quiescence in flexible-bottomed culture plates, Wounds were created in a standardized fashion using a specially designed jig, The jig utilized a 25 gauge needle to reproducibly ablate 16-20% of cells from confluent cell sheets. Wounded and nonwounded cells were labeled with H-3-thymidine at 24, 48, 72, and 96 hr postwounding to measure DNA synthesis. Wounded KF and NDF demonstrated increased H-3-thymidine incorporation compared to nonwounded control cultures, and wounded KF demonstrated significantly higher levels of H-3-thymidine incorporation than wounded NDF both 24 and 48 hr after wounding. A similar trend was seen in cell counts. The wounded KF also showed a statistically greater labeling index quantitated by autoradiography than did wounded NDF, The increased commitment to DNA synthesis in response to wounding in vitro in keloid fibroblasts correlates with pathology seen in vivo. Keloid fibroblasts may have a lower inherent threshold for S phase entry than do normal fibroblasts contributing to the increased proliferation of keloid fibroblasts in response to wounding in vitro. (C) 1996 Academic Press, Inc.